[聚乙二醇与海藻糖联合冻干牛心包的效果]
[Effects of freeze-drying bovine pericardium using a combination of polyethylene glycol and trehalose].
作者信息
Huang Wei, Li Weijie, Liu Baolin
机构信息
Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology, Shanghai 200093, P. R. China.
Shanghai Co-innovation Center for Energy Therapy of Tumors, Shanghai 200093, P. R. China.
出版信息
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2024 Apr 25;41(2):368-375. doi: 10.7507/1001-5515.202311035.
The freeze-drying is a technology that preserves biological samples in a dry state, which is beneficial for storage, transportation, and cost saving. In this study, the bovine pericardium was treated with a freeze-drying protectant composed of polyethylene glycol (PEG) and trehalose (Tre), and then freeze-dried. The results demonstrated that the mechanical properties of the pericardium treated with PEG + 10% w/v Tre were superior to those of the pericardium fixed with glutaraldehyde (GA). The wet state water content of the rehydrated pericardium, determined using the Karl Fischer method, was (74.81 ± 1.44)%, which was comparable to that of the GA-fixed pericardium. The dry state water content was significantly reduced to (8.64 ± 1.52)%, indicating effective dehydration during the freeze-drying process. Differential scanning calorimetry (DSC) testing revealed that the thermal shrinkage temperature of the pericardium was (84.96 ± 0.49) ℃, higher than that of the GA-fixed pericardium (83.14 ± 0.11) ℃, indicating greater thermal stability. Fourier transform infrared spectroscopy (FTIR) results showed no damage to the protein structure during freeze-drying. Hematoxylin and eosin (HE) staining demonstrated that the freeze-drying process reduced pore formation, prevented ice crystal growth, and resulted in a tighter arrangement of tissue fibers. The frozen-dried bovine pericardium was subjected to tests for cell viability and hemolysis rate. The results revealed a cell proliferation rate of (77.87 ± 0.49)%, corresponding to a toxicity grade of 1. Additionally, the hemolysis rate was (0.17 ± 0.02)%, which is below the standard of 5%. These findings indicated that the frozen-dried bovine pericardium exhibited satisfactory performance in terms of cytotoxicity and hemolysis, thus meeting the relevant standards. In summary, the performance of the bovine pericardium treated with PEG + 10% w/v Tre and subjected to freeze-drying could meet the required standards.
冷冻干燥是一种将生物样品保存在干燥状态的技术，这有利于储存、运输和节省成本。在本研究中，牛心包用由聚乙二醇（PEG）和海藻糖（Tre）组成的冷冻干燥保护剂处理，然后进行冷冻干燥。结果表明，用PEG + 10% w/v Tre处理的心包的力学性能优于用戊二醛（GA）固定的心包。使用卡尔费休法测定的复水心包的湿态含水量为（74.81 ± 1.44）%，与GA固定心包的含水量相当。干态含水量显著降低至（8.64 ± 1.52）%，表明在冷冻干燥过程中脱水有效。差示扫描量热法（DSC）测试显示，心包的热收缩温度为（84.96 ± 0.49）℃，高于GA固定心包的热收缩温度（83.14 ± 0.11）℃，表明热稳定性更高。傅里叶变换红外光谱（FTIR）结果表明，冷冻干燥过程中蛋白质结构未受损。苏木精和伊红（HE）染色表明，冷冻干燥过程减少了孔隙形成，防止了冰晶生长，并导致组织纤维排列更紧密。对冷冻干燥的牛心包进行了细胞活力和溶血率测试。结果显示细胞增殖率为（77.87 ± 0.49）%，毒性等级为1级。此外，溶血率为（0.17 ± 0.02）%，低于5%的标准。这些结果表明，冷冻干燥的牛心包在细胞毒性和溶血方面表现出令人满意的性能，从而符合相关标准。总之，用PEG + 10% w/v Tre处理并进行冷冻干燥的牛心包的性能可以达到要求的标准。
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